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Biphasic Affinity Chromatographic Approach for Deep Tyrosine Phosphoproteome Analysis
Deng, Zhenzhen1,2; Dong, Mingming1; Wang, Yan1,2; Dong, Jing1; Li, Shawn S. -C.3,4,5; Zou, Hanfa1; Ye, Mingliang1
Source PublicationANALYTICAL CHEMISTRY
2017-02-21
ISSN0003-2700
DOI10.1021/acs.analchem.6b04288
Volume89Issue:4Pages:2405-2410
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordSH2 DOMAINS ; IDENTIFICATION ; SUPERBINDER ; PROTEOME ; REVEALS
AbstractTyrosine phosphorylation (pTyr) is important for normal physiology and implicated in many human diseases, particularly cancer. Identification of pTyr sites is critical to dissecting signaling pathways and understanding disease pathologies. However, compared with serine/threonine phosphorylation (pSer/pThr), the analysis of pTyr at the proteome level is more challenging due to its low abundance. Here, we developed a biphasic affinity chromatographic approach where Src SH2 superbinder was coupled with NeutrAvidin affinity chromatography, for tyrosine phosphoproteome analysis. With the use of competitive elution agent biotin-pYEEI, this strategy can distinguish high-affinity phosphotyrosyl peptides from low-affinity ones, while the excess competitive agent is readily removed by using NeutrAvidin agarose resin in an integrated tip system. The excellent performance of this system was demonstrated by analyzing tyrosine phosphoproteome of Jurkat cells from which 3,480 unique pTyr sites were identified. The biphasic affinity chromatography method for deep Tyr phosphoproteome analysis is rapid, sensitive, robust, and cost-effective. It is widely applicable to the global analysis of the tyrosine phosphoproteome associated with tyrosine kinase signal transduction.
Language英语
WOS IDWOS:000394724700034
PublisherAMER CHEMICAL SOC
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/169337
Collection中国科学院大连化学物理研究所
Corresponding AuthorYe, Mingliang
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 1000491, Peoples R China
3.Western Univ, Dept Biochem, London, ON N6A 5C1, Canada
4.Western Univ, Dept Oncol, London, ON N6A 5C1, Canada
5.Western Univ, Childrens Hlth Res Inst, Schulich Sch Med & Dent, London, ON N6A 5C1, Canada
Recommended Citation
GB/T 7714
Deng, Zhenzhen,Dong, Mingming,Wang, Yan,et al. Biphasic Affinity Chromatographic Approach for Deep Tyrosine Phosphoproteome Analysis[J]. ANALYTICAL CHEMISTRY,2017,89(4):2405-2410.
APA Deng, Zhenzhen.,Dong, Mingming.,Wang, Yan.,Dong, Jing.,Li, Shawn S. -C..,...&Ye, Mingliang.(2017).Biphasic Affinity Chromatographic Approach for Deep Tyrosine Phosphoproteome Analysis.ANALYTICAL CHEMISTRY,89(4),2405-2410.
MLA Deng, Zhenzhen,et al."Biphasic Affinity Chromatographic Approach for Deep Tyrosine Phosphoproteome Analysis".ANALYTICAL CHEMISTRY 89.4(2017):2405-2410.
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